Medium for plant protoplast culture

Abstract

A culture medium and method for the isolation and culture of plant protoplasts is disclosed which includes in it a quantity of neutral mammalian blood serum. An addition of 1 to 25% serum to protoplast media results in a greater yield of viable protoplast cells, helps to protect the protoplasts in culture, and makes it possible to obtain and maintain corn protoplasts, a heretofore difficult task.

I claim: 
     
       1. In a medium for the in vitro isolation and maintenance of plant protoplast cultures including buffer salts, plant nutrients, sugar and plant hormones in aqueous solution wherein the improvement comprises an addition to the medium of a growth promoting amount of mammalian blood serum which has been heat treated to render said serum essentially free of potentially phytotoxic biological substances therefrom. 
     
     
       2. An improvement as claimed in claim 1 wherein the addition of serum is at least 1% of the medium. 
     
     
       3. An improvement as claimed in claim 2 wherein the addition of serum is no more than 25% of the medium. 
     
     
       4. An improvement as claimed in claim 1 wherein the plant is selected from the group consisting of corn, tomato, carrot and cotton. 
     
     
       5. In a method wherein protoplasts are cultivated in a media and are isolated wherein the improvement comprises adding to the protoplast medium a growth promoting amount of mammalian blood serum which has been heat treated to render said serum essentially free of potentially phytotoxic biological sustances therefrom. 
     
     
       6. A method as claimed in claim 5 wherein the adding step includes adding no less than 1% and no more than 25% serum to the medium. 
     
     
       7. A method as claimed in claim 5 further comprising the step of neutralizing the serum by filtering with activated charcoal and by heat treatment at approximately 57° C. 
     
     
       8. A method as claimed in claim 5 wherein the plant is a cereal plant. 
     
     
       9. A method as claimed in claim 5 wherein the plant is selected from the group consisting of corn, tomato, cotton, and carrot. 
     
     
       10. A method as claimed in claim 5 wherein the serum is neutral whole serum.